Proteomic analysis reveals a PLK1-dependent G2/M degradation program and a role for AKAP2 in coordinating the mitotic cytoskeleton

Cell Rep. 2024 Aug 27;43(8):114510. doi: 10.1016/j.celrep.2024.114510. Epub 2024 Jul 16.

Abstract

Ubiquitination is an essential regulator of cell division. The kinase Polo-like kinase 1 (PLK1) promotes protein degradation at G2/M phase through the E3 ubiquitin ligase Skp1-Cul1-F box (SCF)βTrCP. However, the magnitude to which PLK1 shapes the mitotic proteome is uncharacterized. Combining quantitative proteomics with pharmacologic PLK1 inhibition revealed a widespread, PLK1-dependent program of protein breakdown at G2/M. We validated many PLK1-regulated proteins, including substrates of the cell-cycle E3 SCFCyclin F, demonstrating that PLK1 promotes proteolysis through at least two distinct E3 ligases. We show that the protein-kinase-A-anchoring protein A-kinase anchor protein 2 (AKAP2) is cell-cycle regulated and that its mitotic degradation is dependent on the PLK1/βTrCP signaling axis. Expression of a non-degradable AKAP2 mutant resulted in actin defects and aberrant mitotic spindles, suggesting that AKAP2 degradation coordinates cytoskeletal organization during mitosis. These findings uncover PLK1's far-reaching role in shaping the mitotic proteome post-translationally and have potential implications in malignancies where PLK1 is upregulated.

Keywords: AKAP2; CP: Cell biology; CP: Molecular biology; Cullin RING Ligase; Cyclin F; G2/M; Plk1; cell cycle; mitosis; protein degradation; ubiquitin; βTRCP.

MeSH terms

  • A Kinase Anchor Proteins* / metabolism
  • Cell Cycle Proteins* / metabolism
  • Cytoskeleton / metabolism
  • G2 Phase
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Mitosis*
  • Polo-Like Kinase 1*
  • Protein Serine-Threonine Kinases* / metabolism
  • Proteolysis
  • Proteomics* / methods
  • Proto-Oncogene Proteins* / metabolism

Substances

  • Polo-Like Kinase 1
  • Proto-Oncogene Proteins
  • Cell Cycle Proteins
  • Protein Serine-Threonine Kinases
  • A Kinase Anchor Proteins